CN105259173A - Reagent combination and application thereof, and kit including reagent combination - Google Patents
Reagent combination and application thereof, and kit including reagent combination Download PDFInfo
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- CN105259173A CN105259173A CN201510728075.6A CN201510728075A CN105259173A CN 105259173 A CN105259173 A CN 105259173A CN 201510728075 A CN201510728075 A CN 201510728075A CN 105259173 A CN105259173 A CN 105259173A
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Abstract
The invention relates to the technical field of medical examination, in particular to a reagent combination and an application thereof, and a kit including the reagent combination. The reagent combination comprises a component A and a component B, wherein the component A includes an acid and alkali buffer pair, 2-amino-2-methyl-1-propanol and magnesium ions, and the component B includes disodium 4-nitrophenyl phosphate. The reagent combination can improve bottle open stability of reagents, and has high accuracy, good repeatability, wide linear range and high resistance to interference.
Description
Technical field
The present invention relates to technical field of medical examination, particularly a kind of composite reagent and application thereof and the kit containing this composite reagent.
Background technology
Alkaline phosphatase (ALP) belongs to orthophosphoric ester monohydrolase, is one group of special phosphate.This enzyme is distributed widely in tissue and body fluid, and higher with content in bone, liver, mammary gland, small intestine, kidney, normal human serum alkaline phosphatase produces primarily of osteocyte, and fraction, from liver, is drained through liver and gall.So when ALP produces too much or excretion is obstructed, ALP in blood all can be made to change, ALP measures and is mainly used for diagnosis liver and gall and disease of skeletal system, is occupying lesion and rachitic important indicator in reflection Bile duct obstruction, liver.
Alkaline phosphatase is higher is divided into physiological to raise and pathologic rising, and physiological rising and children's bone puberty, pregnant woman and union phase, the alkaline phosphatase in bone tissue is very active, and measurement result is higher; In disease, property raises and is common in (1) skeletal diseases: as rickets, osteomalacia, bone malignant tumour, malignant metastatic tumor of bone etc.; (2) disease in the liver and gallbladder is as extrahepatic biliary passages obstruction, liver cancer, cirrhosis, Cholestatic hepatitis etc.; (3) other diseases: as hyperparathyroidism.
At present, the detection method of alkaline phosphatase assay kit is mainly with the KINETIC METHOD that 4-Nitrophenyl phosphate sodium (4-NPP) is matrix, the method is international clinical chemistry association recommend method, its principle is for substrate with 4-Nitrophenyl phosphate sodium, the acceptor that 2-amino-2-methyl-1-propanol (AMP) is phosphate acyl, under alkaline environment (about pH10), ALP catalysis 4-NPP is hydrolyzed and produces yellow paranitrophenol, the generating rate of p-nitrophenol is directly proportional to serum activity change of Alkaline phosphatase, increase speed according to 405nm place absorbance thus calculate ALP active unit.
Decaying phenomenon after biochemical reagents uncork is the FAQs in inspection work, the decay of reagent can cause testing result to occur that tendency changes, then be considered as when decay degree exceedes scope or the time limit of reagent factory settings losing efficacy, therefore reagent stability becomes affect reagent and imitates the key factor that phase and testing result make a variation in the daytime.For the hospital of middle and small scale, because its sample size is less, the long period is needed just to be finished one bottle of reagent, therefore higher to the uncork stability requirement of reagent.But it is nearly all undesirable that existing current conventional alkaline phosphatase measures reagent stability, and quality-control product generally can stablize about 13 days, and sample stablizes about 6 days, can not meet clinical detection demand.
Summary of the invention
In view of this, the invention provides a kind of composite reagent and application thereof and the kit containing this composite reagent.Reagent composition provided by the invention reaches and improves reagent stability object, and accuracy is high, reproducible, the range of linearity is wide, antijamming capability is strong.
In order to realize foregoing invention object, the invention provides following technical scheme:
The invention provides a kind of composite reagent of detection of alkaline phosphatase, comprise component A and B component;
Described component A comprise soda acid buffering to, 2-amino-2-methyl-1-propanol, magnesium ion;
Described B component comprises 4-Nitrophenyl phosphate sodium.
In specific embodiments more of the present invention, soda acid described in the described component A of described composite reagent buffering to for sodium carbonate-bicarbonate buffering to or potassium carbonate-bicarbonate cushion right.
In specific embodiments more of the present invention, in the described component A of described composite reagent, also comprise water.
In specific embodiments more of the present invention, the described component A of described composite reagent also comprises a kind of or both the above potpourris in zinc ion, N-beta-hydroxy ethyl-3-acetic acid ethylenediamine, antiseptic.
In specific embodiments more of the present invention, the pH value of the described component A of described composite reagent is 9.0 ~ 11.0.
In specific embodiments more of the present invention, the described B component of described composite reagent also comprises water.
In specific embodiments more of the present invention, the described B component of described composite reagent also comprises antiseptic.
In specific embodiments more of the present invention, described composite reagent in mass parts, comprise following component:
Described component A comprises:
Described B component comprises:
4-NPP0.5 ~ 10 part
90 ~ 99.5 parts, water
The mass ratio of described component A and described B component is 2 ~ 5:1.
In specific embodiments more of the present invention, described composite reagent in mass parts, comprise following component:
Described component A comprises:
Described B component comprises:
4-NPP0.5 ~ 10 part
Antiseptic 0.05 ~ 1.0 part
89 ~ 99.45 parts, water
The mass ratio of described component A and described B component is 2 ~ 5:1.
In specific embodiments more of the present invention, described composite reagent in mass parts, comprise following component:
Described component A comprises:
Described B component comprises:
4-NPP0.5 ~ 10 part
Antiseptic 0.05 ~ 1.0 part
89 ~ 99.45 parts, water
The mass ratio of described component A and described B component is 2 ~ 5:1.
In specific embodiments more of the present invention, described composite reagent in mass parts, comprise following component:
Described component A comprises:
Described B component comprises:
4-NPP0.5 ~ 5 part
Antiseptic 0.05 ~ 0.5 part
94.5 ~ 99.45 parts, water
The mass ratio of described component A and described B component is 2 ~ 5:1.
In specific embodiments more of the present invention, described composite reagent in mass parts, comprise following component:
Described component A comprises:
Described B component comprises:
4-NPP0.5 ~ 5 part
Antiseptic 0.05 ~ 0.5 part
94.5 ~ 99.45 parts, water
The mass ratio of described component A and described B component is 2 ~ 5:1.
In specific embodiments more of the present invention, the mass ratio of described component A and described B component is 4:1.
In specific embodiments more of the present invention, the mass parts of the water in described component A is 70.7 ~ 98.69 parts; The mass parts of the water in described B component is 90 ~ 99.5 parts.
In specific embodiments more of the present invention, described composite reagent in mass percentage, comprise following component:
Described component A comprises:
Described B component comprises:
4-NPP0.5～10％
Water 90 ~ 99.5%
The mass ratio of described component A and described B component is 2 ~ 5:1.
In specific embodiments more of the present invention, described composite reagent in mass percentage, comprise following component:
Described component A comprises:
Described B component comprises:
4-NPP0.5～10％
Antiseptic 0.05 ~ 1.0%
Water 89 ~ 99.45%
The mass ratio of described component A and described B component is 2 ~ 5:1.
In specific embodiments more of the present invention, described composite reagent in mass percentage, comprise following component:
Described component A comprises:
Described B component comprises:
4-NPP0.5～5％
Antiseptic 0.05 ~ 0.5%
Water 94.5 ~ 99.45%
The mass ratio of described component A and described B component is 2 ~ 5:1.
In specific embodiments more of the present invention, described composite reagent in mass percentage, comprise following component:
Described component A comprises:
Described B component comprises: 4-NPP0.5 ~ 10%
Antiseptic 0.05 ~ 1.0%
Water 89 ~ 99.45%
The mass ratio of described component A and described B component is 2 ~ 5:1.
In specific embodiments more of the present invention, described composite reagent in mass percentage, comprise following component:
Described B component comprises: 4-NPP0.5 ~ 5%
Antiseptic 0.05 ~ 0.5%
Water 94.5 ~ 99.45%
The mass ratio of described component A and described B component is 2 ~ 5:1.
In specific embodiments more of the present invention, described acid-base reagent can be HCl or H
2sO
4.
In specific embodiments more of the present invention, described zinc ion is soluble zinc salt, is specifically as follows one or more in zinc sulfate, zinc chloride.
In specific embodiments more of the present invention, described magnesium ion is solubility magnesium salts, is specifically as follows one or more in magnesium chloride, magnesium sulfate, magnesium nitrate.
Present invention also offers the application of described composite reagent in the testing tool preparing alkaline phosphatase.
The kit that present invention also offers a kind of detection of alkaline phosphatase comprises described composite reagent.
In specific embodiments more of the present invention, the mass ratio of sample to be tested and composite reagent provided by the invention is 1:(40-60).
The invention provides a kind of composite reagent of detection of alkaline phosphatase, comprise component A and B component; Described component A comprise soda acid buffering to, 2-amino-2-methyl-1-propanol, magnesium ion; Described B component comprises 4-Nitrophenyl phosphate sodium.
In the present invention, AMP refers to 2-amino-2-methyl-1-propanol, and HEDTA refers to N-beta-hydroxy ethyl-3-acetic acid ethylenediamine, and 4-NPP refers to 4-Nitrophenyl phosphate sodium.Uncork stability test shows: conventional reagents: uncork stability, and quality-control product can stablize 13 days, and sample stablizes 6 days; Reagent in kit of the present invention: uncork stability, quality-control product can stablize 23 days, and sample stablizes 15 days.Other performance index are with now to sell reagent consistent, and accuracy is high, and precision is good, and the range of linearity is wide, and antijamming capability is strong, and performance steady in a long-term stablizes 1 year.
Accuracy test shows, the invention provides to the measurement result of kit and target value deviation meet " alkaline phosphatase assay kit (NPP substrate-AMP buffer method) industry standard " requirement, illustrate that reagent accuracy of the present invention is good.
Replica test result shows, the coefficient of variation CV of seminal plasma fructose detection kit provided by the invention is < 5%, meet " alkaline phosphatase assay kit (NPP substrate-AMP buffer method) industry standard " requirement, and be less than the conventional reagents coefficient of variation, illustrate that kit of the present invention is reproducible.
Range of linearity testing result shows, linear high limit all can accomplish 1000U/L, relative deviation is less than 10%, meet " alkaline phosphatase assay kit (NPP substrate-AMP buffer method) industry standard " requirement, and linear high limit is greater than standard-required 750U/L, illustrate that the kit range of linearity of the present invention is wide, applied widely.
Reagent interference test shows: Vc1.0g/L, haemoglobin 5g/L, chyle TG24mmol/L, cholerythrin 800 μm of ol/L measure all noiseless to reagent, illustrate that kit antijamming capability of the present invention is strong.
Embodiment
The invention discloses a kind of composite reagent and application thereof and the kit containing this composite reagent, those skilled in the art can use for reference present disclosure, and suitable improving technique parameter realizes.Special needs to be pointed out is, all similar replacements and change apparent to those skilled in the art, they are all deemed to be included in the present invention.Method of the present invention and application are described by preferred embodiment, related personnel obviously can not depart from content of the present invention, spirit and scope methods and applications as herein described are changed or suitably change with combination, realize and apply the technology of the present invention.
A kind of composite reagent provided by the invention and application thereof and all can be buied by market containing agents useful for same and raw material in the kit of this composite reagent.Below in conjunction with embodiment, set forth the present invention further:
Embodiment 1: alkaline phosphatase assay kit reagent is prepared
B component is prepared: 4-NPP0.5%
Water 99.5%
Wherein, the pH of component A is 9.0; By volume, component A: B component=2:1.
Embodiment 2: alkaline phosphatase assay kit reagent is prepared
B component is prepared: 4-NPP10%
Water 90%
Wherein, the pH of component A is 11.0; By volume, component A: B component=3:1.
Embodiment 3: alkaline phosphatase assay kit reagent is prepared
B component is prepared: 4-NPP0.5%
Antiseptic 0.05%
Water 99.45%
Wherein, the pH of component A is 9.5; By volume, component A: B component=4:1.
Embodiment 4: alkaline phosphatase assay kit reagent is prepared
B component is prepared: 4-NPP5.0%
Antiseptic 0.5%
Water 94.5%
Wherein, the pH of component A is 10.5; By volume, component A: B component=5:1.
Embodiment 5: alkaline phosphatase assay kit reagent is prepared
B component is prepared: 4-NPP2.5%
Antiseptic 0.1%
Water 97.4%
Wherein, the pH of component A is 10.0, by volume, and component A: B component=4:1.
Embodiment 6: alkaline phosphatase assay kit reagent is prepared
B component is prepared: 4-NPP2.5%
Antiseptic 0.1%
Water 97.4%
Wherein, the pH of component A is 10.0; By volume, component A: B component=4:1.
Embodiment 7: alkaline phosphatase assay kit reagent is prepared
B component is prepared: 4-NPP2.5%
Antiseptic 0.1%
Water 97.4%
Wherein, the pH of component A is 10.0; By volume, component A: B component=2:1.
Embodiment 8: alkaline phosphatase assay kit reagent is prepared
B component is prepared: 4-NPP2.5%
Antiseptic 0.1%
Water 97.4%
Wherein, the pH of component A is 10.0; By volume, component A: B component=5:1.
Comparative example 1: alkaline phosphatase assay kit reagent is prepared
B component is prepared: 4-NPP2.5%
Antiseptic 0.1%
Water 97.4%
Wherein, the pH of component A is 10.0; By volume, component A: B component=4:1.
Comparative example 2: alkaline phosphatase assay kit reagent is prepared
B component is prepared: 4-NPP2.5%
Water 97.5%
Wherein, the pH of component A is 10.0; By volume, component A: B component=4:1.
Embodiment 11: alkaline phosphatase assay kit test method
1. instrument: there is automatic clinical chemistry analyzer or semi-automatic biochemical analyzer that wavelength is 405nm.
2. optimum configurations and operation
Method: rate method predominant wavelength: 405nm commplementary wave length: 505nm
Temperature of reaction: 37 DEG C of reaction time: 10min
Sample measures concrete operation method: sample first with reagent component A37 DEG C of incubation 5 minutes, then add B component reagent 37 DEG C of incubations after 1 minute, measure initial absorbance, then Accurate Determining average minute clock absorbance rate of change (Δ A/min).
Wherein, by volume, component A: B component=2 ~ 5:1, (component A+ B component): sample=40 ~ 60:1.
In formula:
Δ A
u/ min-sample absorbance rate of change; Δ A
c/ min-calibration object absorbance rate of change; C
c--calibration object concentration.
Operation steps:
1) set on instrument according to reagent parameter, reagent is put into instrument, select 2 calibrations, click start button;
2) quality-control product and sample measure: quality-control product or sample are put on instrument sample rack, at instrument working interface editing sample test event and test number, click start button.
Embodiment 12: alkaline phosphatase assay kit uncork stability experiment
1. get biochemistry quality control product level 1 and 2, after volume adds purified water redissolution according to the rules, packing is stored in-20 DEG C, for reagent stability observing.
2. get 10 routine clinical samples, small size packing is stored in-20 DEG C, for reagent stability observing.
3. after the alkaline phosphatase assay kit that Example 1 to 5 provides is calibrated according to the method described above, measure quality-control product and 10 routine clinical samples, reagent opening is put on instrument, measures quality-control product and clinical sample, with deviation 10% for control line, observe reagent stability.
The uncork data of the kit measurement quality-control product that embodiment 1 to 8 provides are in table 1 ~ table 8.
The uncork data of the kit measurement quality-control product that table 1 embodiment 1 provides
The uncork data of the kit measurement quality-control product that table 2 embodiment 2 provides
The uncork data of the kit measurement quality-control product that table 3 embodiment 3 provides
The uncork data of the kit measurement quality-control product that table 4 embodiment 4 provides
The uncork data of the kit measurement quality-control product that table 5 embodiment 5 provides
The uncork data of the kit measurement quality-control product that table 6 embodiment 6 provides
The uncork data of the kit measurement quality-control product that table 7 embodiment 7 provides
The uncork data of the kit measurement quality-control product that table 8 embodiment 8 provides
The uncork data of the kit measurement clinical sample that embodiment 1 to 8 provides are in table 9 ~ table 16.
The uncork data of the kit measurement clinical sample that table 9 embodiment 1 provides
The uncork data of the kit measurement clinical sample that table 10 embodiment 2 provides
The uncork data of the kit measurement clinical sample that table 11 embodiment 3 provides
The uncork data of the kit measurement clinical sample that table 12 embodiment 4 provides
The uncork data of the kit measurement clinical sample that table 13 embodiment 5 provides
The uncork data of the kit measurement clinical sample that table 14 embodiment 6 provides
The uncork data of the kit measurement clinical sample that table 15 embodiment 7 provides
The uncork data of the kit measurement clinical sample that table 16 embodiment 8 provides
Alkaline phosphatase assay kit carries out above-mentioned uncork stability test, alkaline phosphatase assay kit results prepared by result and embodiment 1 ~ 8 is close, without significant difference (P > 0.05), quality-control product can stablize 23 days, and clinical sample stablizes 15 days.
Comparative example 1 kit detects quality-control product and the results are shown in Table 17:
Table 17 comparative example 1 kit detects quality-control product result
| Uncork number of days | Quality-control product 1 | Deviation | Quality-control product 2 | Deviation |
| 0 | 116 | / | 410 | / |
| 1 | 119.5 | 3.02％ | 413 | 0.73％ |
| 2 | 116 | 0.00％ | 401.5 | -2.07％ |
| 5 | 108 | -6.90％ | 400 | -2.44％ |
| 6 | 111 | -4.31％ | 402.5 | -1.83％ |
| 7 | 109 | -6.03％ | 405 | -1.22％ |
| 9 | 108 | -6.90％ | 399.5 | -2.56％ |
| 10 | 108.5 | -6.47％ | 385 | -6.10％ |
| 12 | 106 | -8.62％ | 388 | -5.37％ |
| 14 | 105 | -9.48％ | 385 | -6.10％ |
| 15 | 103 | -11.21％ | 375 | -8.54％ |
| 19 | 102 | -12.07％ | 368 | -10.24％ |
| 23 | 99 | -14.66％ | 360 | -12.20％ |
Comparative example 1 kit detects clinical sample and the results are shown in Table 18.
Table 18 comparative example 1 kit detects clinical sample result
The kit that comparative example 2 provides detects quality-control product and the results are shown in Table 19:
The kit that table 19 comparative example 2 provides detects quality-control product result
| Uncork number of days | Quality-control product 1 | Deviation | Quality-control product 2 | Deviation |
| 0 | 112 | / | 405 | / |
| 1 | 118 | 5.36％ | 412 | 1.73％ |
| 2 | 113 | 0.89％ | 403 | -0.49％ |
| 5 | 108 | -3.57％ | 399 | -1.48％ |
| 6 | 106 | -5.36％ | 401.5 | -0.86％ |
| 7 | 109 | -2.68％ | 405 | 0.00％ |
| 11 | 103 | -8.04％ | 388 | -4.20％ |
| 13 | 105 | -6.25％ | 391 | -3.46％ |
| 14 | 106 | -5.36％ | 385 | -4.94％ |
| 15 | 103 | -8.04％ | 382 | -5.68％ |
| 16 | 102 | -8.93％ | 371 | -8.40％ |
| 19 | 100 | -10.71％ | 367 | -9.38％ |
| 23 | 98 | -12.50％ | 360 | -11.11％ |
The kit that comparative example 2 provides detects clinical sample and the results are shown in Table 20:
The kit that table 20 comparative example 2 provides detects clinical sample result
Conventional reagents box detects quality-control product and the results are shown in Table 21:
| Uncork number of days | Quality-control product 1 | Deviation | Quality-control product 2 | Deviation |
| 0 | 114 | / | 456 | / |
| 1 | 122.5 | 7.46％ | 475.5 | 4.28％ |
| 2 | 114 | 0.00％ | 457 | 0.22％ |
| 5 | 112 | -1.75％ | 447 | -1.97％ |
| 6 | 111 | -2.63％ | 446.5 | -2.08％ |
| 7 | 109 | -4.39％ | 444.5 | -2.52％ |
| 11 | 106.5 | -6.58％ | 432.5 | -5.15％ |
| 13 | 104.5 | -8.33％ | 421.5 | -7.57％ |
| 14 | 101 | -11.40％ | 412 | -9.65％ |
| 15 | 102 | -10.09％ | 412 | -9.65％ |
| 16 | 100 | -12.28％ | 400.5 | -12.17％ |
| 19 | 94 | -17.54％ | 383 | -16.01％ |
| 23 | 82 | -28.07％ | 331 | -27.41％ |
Conventional reagents box detects clinical sample and the results are shown in Table 22:
Conclusion: alkaline phosphatase assay kit uncork stability of the present invention is all better than conventional reagents, and quality-control product can stablize 23 days, and clinical sample stablizes 15 days; The Quality Control of conventional reagents uncork stability stablizes 13 days, and sample can only stablize 6 days.
Embodiment 13: other performance evaluations of embodiment 5 alkaline phosphatase assay kit
1. accuracy: measure alkaline phosphatase quality-control product, replication 3 times, calculates measured value and target value deviation.In table 23:
Table 23 accuracy estimating result
Conclusion: found out by above result, measurement result and target value deviation < 10%, meet " alkaline phosphatase assay kit (NPP
Substrate-AMP buffer method) industry standard " requirement, illustrate that reagent accuracy of the present invention is good.
2. repeated experiment: using routine clinical sample as testing sample, the seminal plasma fructose detection kit replication obtained by conventional reagents and the embodiment of the present invention 5 10 times, calculates coefficient of variation CV%.In table 24:
Table 24 reproducibility result
Conclusion: from above result, the coefficient of variation CV of the seminal plasma fructose detection kit that the embodiment of the present invention 5 provides is 0.74%, 0.29%, < 5%, meet " alkaline phosphatase assay kit (NPP substrate-AMP buffer method) industry standard " requirement, and be less than the conventional reagents coefficient of variation, illustrate that kit of the present invention is reproducible.
3. the range of linearity detects: get alkaline phosphatase concentration close to 1000U/L serum high level, be diluted to 5 different concentration (xi), reagent in the kit provided by the embodiment of the present invention 5 according to each concentration determination of detection method 3 times, calculating mean value (yi).Being independent variable with xi, is that dependent variable obtains equation of linear regression with yi, calculates linear regression correlation coefficient r; Xi is substituted into equation of linear regression, calculates the estimated value of yi and the deviation of yi and estimated value.In table 25:
Table 25 range of linearity testing result
Conclusion: linearly dependent coefficient r is 1.00, is greater than 0.99; Within the scope of 25-100U/L, absolute deviation is 1.07U/L, be less than 10U/L, within the scope of 100-1000U/L, relative deviation is less than 10%, meets " alkaline phosphatase assay kit (NPP substrate-AMP buffer method) industry standard " requirement, and linear high limit is greater than standard-required 750U/L, illustrate that the kit range of linearity of the present invention is wide, applied widely.
4. reagent interference experiment: disturb concentration (g/L) with basal serum preparation Vc: 0.2,0.4,0.6,0.8,1.0; Haemoglobin (g/L): 1,2,3,4,5; Chyle (TGmmol/L): 1000,2000,4000,5000,6000; Cholerythrin (μm ol/L): 100,200,400,600,800 a series of analyzing samples, the kit Simultaneously test provided by the embodiment of the present invention 5 is not containing basal serum and above a series of analyzing samples of interfering material, the relative deviation of computational analysis sample and basal serum, with 10% for interference control line, investigate kit antijamming capability.In table 26:
Table 26 reagent interference test result
Conclusion: the visible Vc1.0g/L of above result, haemoglobin 5g/L, chyle TG24mmol/L, cholerythrin 800 μm of ol/L measure all noiseless to reagent, illustrate that kit antijamming capability of the present invention is strong.
Reagent in the kit provided embodiment 1 ~ 4, embodiment 6 ~ 8 by the method for embodiment 14 carries out above-mentioned performance evaluation, result:
1. accuracy, embodiment 1 ~ 4, the equal <5% of embodiment 6 ~ 8 reagent RE/%, experimental result is good;
2. repeatability, embodiment 1 ~ 4, embodiment 6 ~ 8 reagent are respectively 0.81%, 0.72%, 0.83%, 0.78%, 0.79%, 0.8%, 0.81%, equal <5%, and experimental result is all good;
3. linear, the linear high limit of embodiment 1 ~ 4, embodiment 6 ~ 8 reagent all can accomplish 1000U/L, and experimental result is good;
4. disturb, embodiment 1 ~ 4, embodiment 6 ~ 8 reagent, Vc1.0g/L, haemoglobin 5g/L, chyle TG24mmol/L, cholerythrin 800 μm of ol/L measure all noiseless to reagent, illustrate that reagent antijamming capability is strong.
The corresponding test findings of the seminal plasma fructose detection kit that the test findings of above-described embodiment 1 ~ 4, embodiment 6 ~ 8 is prepared to embodiment 5 is close, without significant difference (P > 0.05).Show: the accuracy of kit provided by the invention, repeatability, linear, antijamming capability is strong.
The above is only the preferred embodiment of the present invention; it should be pointed out that for those skilled in the art, under the premise without departing from the principles of the invention; can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.
Claims (11)
1. a composite reagent for detection of alkaline phosphatase, is characterized in that, comprises component A and B component;
Described component A comprise soda acid buffering to, 2-amino-2-methyl-1-propanol, magnesium ion;
Described B component comprises 4-Nitrophenyl phosphate sodium.
2. composite reagent according to claim 1, is characterized in that, soda acid described in described component A buffering to for sodium carbonate-bicarbonate buffering to or potassium carbonate-bicarbonate cushion right.
3. composite reagent according to claim 1 and 2, is characterized in that, described component A also comprises a kind of or both the above potpourris in zinc ion, N-beta-hydroxy ethyl-3-acetic acid ethylenediamine, antiseptic, water.
4. the composite reagent according to any one of claims 1 to 3, is characterized in that, the pH value of described component A is 9.0 ~ 11.0.
5. the composite reagent according to any one of Claims 1-4, is characterized in that, described B component also comprises water and/or antiseptic.
6. composite reagent according to claim 5, is characterized in that, in described component A, magnesium ion is solubility magnesium salts, is preferably at least one in magnesium chloride, magnesium sulfate, magnesium nitrate; In described component A, zinc ion is soluble zinc salt, is preferably at least one in zinc sulfate, zinc chloride.
7. the composite reagent according to any one of claim 1 to 6, is characterized in that, in mass parts, comprises following component:
Described component A comprises:
Described B component comprises:
0.5 ~ 10 part, 4-Nitrophenyl phosphate sodium
90 ~ 99.5 parts, water
The mass ratio of described component A and described B component is 2 ~ 5:1.
8. the composite reagent according to any one of claim 1 to 7, is characterized in that, in mass parts, comprises following component:
Described component A comprises:
Described B component comprises:
0.5 ~ 10 part, 4-Nitrophenyl phosphate sodium
Antiseptic 0.05 ~ 1.0 part
89 ~ 99.45 parts, water
The mass ratio of described component A and described B component is 2 ~ 5:1.
9. the composite reagent according to any one of claim 1 to 8, is characterized in that, in mass parts, comprises following component:
Described component A comprises:
Described B component comprises:
0.5 ~ 10 part, 4-Nitrophenyl phosphate sodium
Antiseptic 0.05 ~ 1.0 part
89 ~ 99.45 parts, water
The mass ratio of described component A and described B component is 2 ~ 5:1.
10. the application of the composite reagent according to any one of claim 1 to 9 in the testing tool preparing alkaline phosphatase.
The kit of 11. 1 kinds of detection of alkaline phosphatase, is characterized in that, comprises the composite reagent as described in any one of claim 1 to 9.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510728075.6A CN105259173B (en) | 2015-11-02 | 2015-11-02 | A kind of composite reagent and its application and the kit containing the composite reagent |
| CN201710755835.1A CN107643257B (en) | 2015-11-02 | 2015-11-02 | Combined reagent, application thereof and kit containing combined reagent |
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|---|---|---|---|
| CN201510728075.6A CN105259173B (en) | 2015-11-02 | 2015-11-02 | A kind of composite reagent and its application and the kit containing the composite reagent |
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| CN201710755835.1A Division CN107643257B (en) | 2015-11-02 | 2015-11-02 | Combined reagent, application thereof and kit containing combined reagent |
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| CN105259173A true CN105259173A (en) | 2016-01-20 |
| CN105259173B CN105259173B (en) | 2018-01-30 |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111876465A (en) * | 2020-07-30 | 2020-11-03 | 武汉生之源生物科技股份有限公司 | High-stability ALP (alpha-amyloid peptide) determination kit and preparation method and application thereof |
| WO2022006373A1 (en) * | 2020-07-01 | 2022-01-06 | Vision Diagnostics, Inc. | Assay and method for measuring alkaline phosphatase activity in urine as an index of tissue zing deficiency |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1329672A (en) * | 1998-10-08 | 2002-01-02 | 罗赫诊断器材股份有限公司 | Method for determining alkaline phosphatase and eliminating haemoglobin disturbances |
| CN1492936A (en) * | 2001-04-17 | 2004-04-28 | 国际试药株式会社 | Method of assaying biological component |
| CN102297962A (en) * | 2011-05-23 | 2011-12-28 | 董理 | Kit for detecting alkaline phosphatase |
| CN103276051A (en) * | 2013-05-24 | 2013-09-04 | 宁波美康生物科技股份有限公司 | Alkaline phosphatase detection reagent |
| CN103675282A (en) * | 2013-11-27 | 2014-03-26 | 新昌县美迪生物科技有限公司 | Quantitative bone alkaline phosphatase detection kit and application thereof |
| CN104263831A (en) * | 2014-09-28 | 2015-01-07 | 南京诺唯赞生物科技有限公司 | Method for simply, conveniently and quickly determining activity of alkaline phosphatase |
| CN104990912A (en) * | 2015-06-26 | 2015-10-21 | 苏州浩欧博生物医药有限公司 | Enzyme-promoting chemiluminiscence substrate using alkaline phosphatase |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4555484A (en) * | 1983-07-25 | 1985-11-26 | Eastman Kodak Company | Analytical element and method for alkaline phosphatase assay |
-
2015
- 2015-11-02 CN CN201510728075.6A patent/CN105259173B/en active Active
- 2015-11-02 CN CN201710755835.1A patent/CN107643257B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1329672A (en) * | 1998-10-08 | 2002-01-02 | 罗赫诊断器材股份有限公司 | Method for determining alkaline phosphatase and eliminating haemoglobin disturbances |
| CN1492936A (en) * | 2001-04-17 | 2004-04-28 | 国际试药株式会社 | Method of assaying biological component |
| CN102297962A (en) * | 2011-05-23 | 2011-12-28 | 董理 | Kit for detecting alkaline phosphatase |
| CN103276051A (en) * | 2013-05-24 | 2013-09-04 | 宁波美康生物科技股份有限公司 | Alkaline phosphatase detection reagent |
| CN103675282A (en) * | 2013-11-27 | 2014-03-26 | 新昌县美迪生物科技有限公司 | Quantitative bone alkaline phosphatase detection kit and application thereof |
| CN104263831A (en) * | 2014-09-28 | 2015-01-07 | 南京诺唯赞生物科技有限公司 | Method for simply, conveniently and quickly determining activity of alkaline phosphatase |
| CN104990912A (en) * | 2015-06-26 | 2015-10-21 | 苏州浩欧博生物医药有限公司 | Enzyme-promoting chemiluminiscence substrate using alkaline phosphatase |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2022006373A1 (en) * | 2020-07-01 | 2022-01-06 | Vision Diagnostics, Inc. | Assay and method for measuring alkaline phosphatase activity in urine as an index of tissue zing deficiency |
| CN111876465A (en) * | 2020-07-30 | 2020-11-03 | 武汉生之源生物科技股份有限公司 | High-stability ALP (alpha-amyloid peptide) determination kit and preparation method and application thereof |
| CN111876465B (en) * | 2020-07-30 | 2022-11-25 | 武汉生之源生物科技股份有限公司 | High-stability ALP (alpha-amyloid peptide) determination kit and preparation method and application thereof |
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| CN107643257B (en) | 2021-01-01 |
| CN105259173B (en) | 2018-01-30 |
| CN107643257A (en) | 2018-01-30 |
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